Bacillus species LU4 is an effective producer of thermostable site-specific endonuclease BspLU4I,an isoschizomer of AvaI |
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Authors: | Svadbina I V Zheleznyakova E N Zheleznaya L A Matvienko N I |
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Affiliation: | (1) Russian Academy of Sciences, Institute of Theoretical and Experimental Biophysics, Pushchino, Moscow Region, 142290, Russia;(2) Russian Academy of Sciences, Institute of Protein Research, Pushchino, Moscow Region, 142290, Russia |
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Abstract: | The site-specific endonuclease BspLU4I was discovered in the thermophilic Bacillus species LU4 strain and purified to functionally pure state by chromatography on blue agarose, hydroxyapatite HTP, and heparin-Sepharose columns. Analysis of cleavage patterns of different DNAs with known nucleotide sequences demonstrated that the enzyme recognizes the CPyCGPuG site on the DNA. Cleavage points in the sequence were determined with the elongated primer method. It was shown that the endonuclease is an isoschizomer of AvaI. The final yield of the enzyme is 2.25·106 units per g wet biomass. |
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Keywords: | restriction endonucleases isoschizomers AvaI DNA size markers |
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